Cell Lysate

HeLa cells grown in tissue culture and stained with antibody to vimentin, a cytoplasmic filament protein (red); staining of nuclear pore complex protein (green).

Cell lysates are useful in Western Blot applications as a positive control for many antibodies. Most cell lysates are prepared by homogenization in modified RIPA buffer (150mM sodium chloride, 50mM Tris-HCl, pH 7.4, 1mM ethylenediaminetetraacetic acid, 1mM phenylmethylsulfonyl flouride, 1% Triton X-100, 1% sodium deoxycholic acid, 0.1% sodium dodecylsulfate, 5ug/ml of aprotinin, 5ug/ml of leupeptin). Cell debris is removed by centrifugation. Protein concentration is determined with Bio-Rad protein assay. The cell lysate is boiled for 5 minutes in 1 x SDS sample buffer (50mM Tris-HCl pH 6.8, 12.5% glycerol, 1% sodium dodecylsulfate, 0.01% bromophenol blue) containing 5% b-mercaptoethanol.

Selected Cell Lysate Products